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. 2017 Jun 15;28(12):1676–1687. doi: 10.1091/mbc.E16-12-0863

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© 2017 Bottanelli et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — The majority of ARF1 tubules are decorated by clathrin clusters. ARF1^EN-Halo (green) cells were electroporated with a plasmid encoding for SNAP-CLC (magenta). (a–c) Cells were labeled with 590-CA and SiR-BG for two-color live-cell STED imaging on a custom-built microscope. (b, c) Examples of Golgi-derived tubules are highlighted by arrows. (d) A two-dimensional Lorentzian function was fitted to images of clathrin clusters, and FWHM of the fitted functions is represented in histograms. The average size is 99 ± 18 nm for both clathrin clusters on the tubules and on the Golgi (50 clusters). (e) A line profile along the tubule in b shows that there is no enrichment of ARF1 in the clathrin clusters. (f) Quantification of the number of clathrin-positive tubules shows that 73 ± 13% of the tubules are decorated by SNAP-CLC clusters. All STED images were deconvolved; the line profile represents raw image data. Error bars represent SD. Scale bars, 5 μm (a), 1 μm (b, c).